Cancer is a leading cause of death in the United States with over 1,000,000 new cases being diagnosed every year. As a result of the high sensitivities and specificities that are required to justify large-scale population screening, only very few single protein biomarkers are routinely used today in the clinical setting. It is of critical priority to develop novel technologies that will enable the rapid detection of a plethora of biomarkers relevant to early diagnosis, prognosis, staging and treatment response. ? ? The long-term objective of this research is to combine the emerging technology of microfluidics with state-of-the-art mass spectrometry (MS) detection to enhance our capacity for analyzing molecular structure and function in biological systems. This application capitalizes on the distinguishing capabilities of microfluidic architectures that enable process integration, multiplexing, fast and high-throughput processing of minute amounts of sample, and the power of MS detection that provides the sensitivity, specificity and resolving power necessary for unambiguous detection of trace level components. ? ? Specific Aim 1: Develop a compact, low-cost and disposable microfluidic analysis platform with matrix assisted laser desorption ionization (MALDI)-MS/MS detection for high-throughput proteomics that will enable the study of protein co-expression patterns and biomarker discovery. The microfluidic device will carry out parallel liquid chromatography (LC) separations and will integrate a novel microchip-MS interface to enable sensitive MALDI-MS/MS detection directly from the chip. ? ? Specific Aim 2: Demonstrate the effectiveness of the microfluidic MALDI-MS/MS platform for the detection of multiple cancer biomarkers in cellular extracts. Cellular fractions from the MCF7 breast cancer cell line will be analyzed for target proteins that are involved in essential cellular processes driving cancer on-set and development (cell proliferation, cell cycle regulation, DNA repair, apoptosis and invasion/metastasis). ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21CA126669-01A1
Application #
7387166
Study Section
Special Emphasis Panel (ZCA1-SRLB-Q (O1))
Program Officer
Kagan, Jacob
Project Start
2008-06-16
Project End
2010-05-31
Budget Start
2008-06-16
Budget End
2009-05-31
Support Year
1
Fiscal Year
2008
Total Cost
$173,020
Indirect Cost
Name
Virginia Polytechnic Institute and State University
Department
Type
Organized Research Units
DUNS #
003137015
City
Blacksburg
State
VA
Country
United States
Zip Code
24061
Lazar, Iulia M; Hoeschele, Ina; de Morais, Juliana et al. (2017) Cell Cycle Model System for Advancing Cancer Biomarker Research. Sci Rep 7:17989
Tenga, Milagros J; Lazar, Iulia M (2014) Proteomic study reveals a functional network of cancer markers in the G1-Stage of the breast cancer cell cycle. BMC Cancer 14:710
Lazar, Iulia M; Kabulski, Jarod L (2013) Microfluidic LC device with orthogonal sample extraction for on-chip MALDI-MS detection. Lab Chip 13:2055-65
Tenga, Milagros J; Lazar, Iulia M (2013) Proteomic snapshot of breast cancer cell cycle: G1/S transition point. Proteomics 13:48-60
Cortes, Diego F; Kabulski, Jarod L; Lazar, Alexandru C et al. (2011) Recent advances in the MS analysis of glycoproteins: Capillary and microfluidic workflows. Electrophoresis 32:14-29
Tenga, Milagros J; Lazar, Iulia M (2011) Impact of peptide modifications on the isobaric tags for relative and absolute quantitation method accuracy. Anal Chem 83:701-7
Lazar, Iulia M; Lazar, Alexandru C; Cortes, Diego F et al. (2011) Recent advances in the MS analysis of glycoproteins: Theoretical considerations. Electrophoresis 32:3-13
Armenta, Jenny M; Perez, Milagros; Yang, Xu et al. (2010) Fast proteomic protocol for biomarker fingerprinting in cancerous cells. J Chromatogr A 1217:2862-70
Yang, Xu; Lazar, Iulia M (2009) MRM screening/biomarker discovery with linear ion trap MS: a library of human cancer-specific peptides. BMC Cancer 9:96